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Objective DNA methylation, as an epigenetic alteration, plays an essential role in the development of atherosclerosis and venous thrombosis. E-cadherin, a tumor suppressor gene and adhesion molecule, has a crucial function in platelet aggregation and hemostasis. P16, a cell cycle regulator, is involved in venous thrombosis. The aim of this study is to evaluate the DNA methylation patterns and expression levels of the E-cadherin and P16 genes in venous thromboembolism (VTE). Method Peripheral blood samples were collected from 32 patients, including those with deep vein thrombosis (DVT, n = 15), pulmonary embolism (PE, n = 8), DVT with PE (n = 4), intestinal thrombosis (IT, n = 3), and cerebral venous sinus thrombosis (CVST, n = 2), as well as from 10 healthy individuals. The DNA methylation patterns and gene expression levels of E-cadherin and P16 were analyzed using methylation-specific PCR (MSP) and Real-Time PCR, respectively. Results The promoter of the CDH1 gene was partially methylated in 84.4% of thrombotic patients and unmethylated in 15.6% ( P = 0.183). A significantly higher expression level of CDH1 was observed in the patients compared to the controls ( P = 0.001). The P16 gene promoter were unmethylated in all control and patient specimens. Compared to normal subjects, the expression level of the P16 was significantly increased in patients ( P = 0.000). Conclusion Our results indicated that DNA methylation is not the main gene expression regulatory mechanism for E-cadherin and P16 genes in thrombosis. Higher transcription levels of CDH1 and P16 in thrombotic patients may show their crucial roles in the pathogenesis of VTE.

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Objective Chronic myeloid leukemia (CML) is a myeloproliferative malignancy with different stages. Aberrant epigeneticmodifications, such as DNA methylation, have been introduced as a signature for diverse cancers which also plays acrucial role in CML pathogenesis and development. Suppressor with morphogenetic effect on genitalia (SMG1) generecently has been brought to the spotlight as a potent tumor suppressor gene that can be suppressed by tumors forfurther progress. The present study aims to investigate SMG1 status in CML patients. Materials and Methods In this case-control study, peripheral blood from 30 patients with different phases of CML [newcase (N)=10, complete molecular remission (CMR)=10, blastic phase (BP)=10] and 10 healthy subjects were collected.Methylation status and expression level of SMG1 gene promoter was assessed by methylation-specific polymerasechain reaction (MSP) and quantitative reverse-transcription PCR, respectively. Results MSP results of SMG1 gene promotor in the new case group were methylated (60% methylated, 30%hemimethylated and 10% unmethylated). All CMR and control group patients were unmethylated in the SMG1 genepromoter. In the BP group, methylated SMG1 promoter was seen (50% of patients had a methylated status and 50%had hemimethylated status). In comparison with the healthy subjects, expression level of SMG1 in the new case groupwas decreased (P

Abstract Background and aim: Pain is known as the greatest complication of cancer and cancer-related therapies. Therefore, control of pain has been considered with pharmaceutical and non-pharmaceutical methods. This study was performed to survey of mental and psychological activities in control of pain in patients with breast cancer. Methods and materials: 166 patients with breast cancer were selected randomly in a specialist hospital in Tehran, Iran and the questionnaire, and the consent letter was completed by them. Patients were categorized into three groups; first group includes patients with great spirits and hope for complete recovery. Second group includes patients with major depression, and third group includes patients with moderate spirits. Most data in this study were for the primary group. Results: According to data, in the deviation group, no acceptable mean of pain was obtained. In the hypothesis group, mean of pain was more passable. In the group which patients were conversed with own, mean of pain was more than previous two groups. The lowest mean of pain was related to the negative hypothesis group, and the highest mean of pain was related to patients who were prayed and hoped. Conclusion: The obtained results showed that deviation and activity of mental are good way to control and reduce of pain. This method is without side effects, inexpensive and accessible, which reduces of pain in patients. This study suggests that use of mental activity is a non-pharmacological treatment to reduce of pain in outpatient and hospitalized patients.

Background: Breast cancer (BC) is the most common malignancy in women worldwide. The methylation status of MyoD1, a tumor suppressor gene, is enrolled in various cancers, i.e., BC. Various studies showed the impact of MyoD1 epigenetic dysregulation in BC. This study aimed to investigate the methylation status and expression level of MyoD1 in BC patients and its association with the expression of DNMT1. Materials and Methods: This case-control study was conducted on 30 cases (pathology-confirmed ductal carcinoma) and 18 controls (fibroadenoma and fibrocystic masses), referred to Velayat Hospital, Qazvin, Iran. The expression of the MyoD1 and DNMT1 and the promoter methylation of the MyoD1 were evaluated in tissue blocks of BC patient masses using qRT-PCR and MS-PCR assays, respectively. SPSS 24.0 was used to analyze the data. Results: The MyoD1 promoter is hypermethylated in BC patients compared to controls (p =0.001). The expression level of MyoD1 in BC patients was significantly reduced compared to controls (fold change =0.13, p =0.042). In addition, in BC patients, the reduced expression level of MyoD1 was significantly associated with methylation of the MyoD1 promoter (p =0.001). There is no significant difference between the expression level of DNMT1 in BC patients and controls (p =0.197). A significant association is found between the expression of DNMT1 and the methylation status of the MyoD1 promoter (p =0.038). Discussion: The expression level of MyoD1 is affected by the methylation status of the promoter of this gene. Moreover, the expression level and methylation status of MyoD1 are correlated with clinical parameters.

Objectives: Epigenetic alterations, including any change in DNA methylation pattern, could be the missing link of understanding radiation-induced genomic instability. Dapper, Dishevelled-associated antagonist of β-catenin homolog 2 (DACT2) is a tumor suppressor gene regulating Wnt/β-catenin. In hepatocellular carcinoma (HCC), DACT2 is hypermethylated, while methylation status of its promoter regulates the corresponding expression. Radionuclides have been used to reduce proliferation and induce apoptosis in cancerous cells. Epigenetic impact of radionuclides as therapeutic agents for treatment of HCC is still unknown. The aim of this study was to evaluate epigenetic impact of 188Rhenium perrhenate (188ReO4) on HCC cells.Material and Methods: In this in vitro experimental study, HepG2 and Huh7 cells were treated with 188ReO4, receiving 55 and 73 Mega Becquerel (MBq) exposures, respectively. For cell viability measurement, live/dead staining was carried out 18, 24, and 48 hours post-exposure. mRNA expression level of β-Catenin, Wnt1, DNMT1, DACT2 and WIF-1 genes were quantified by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Then, possible regulatory impact of DACT2 upregulation was investigated through evaluating methylation-specific PCR (MS-PCR).Results: Results showed that viability of both cells was reduced after treatment with 188ReO4 at three time points postexposure compared to the control groups. The qRT-PCR results showed that DACT2 mRNA level was significantly increased at 24, and 48 hours post-exposure in HepG2 cells compared to the control group, while, no significant change was observed in Huh7 cells. Methylation pattern of DACT2 promoter remained unchanged in HepG2 and Huh7 cells.Conclusion: Treatment with 188ReO4 reduced viability of HepG2 and Huh7 cells. Although DACT2 expression was increased after 188ReO4 exposure in HepG2 cells, methylation pattern of its promoter remained unchanged. This study assessed impacts of the 188 ReO4 β-irradiation on expression and induction of DACT2 epigenetic aberrations as well as the correlation of this agent with viability of cells.

Severe combined immunodeficiency (SCID) is classified as a primary immunodeficiency, which is characterized by impaired T-lymphocytes differentiation. IL2RG, IL7Ralpha, JAK3, ADA, RAG1/RAG2, and DCLE1C (Artemis) are the most defective genes in SCID. The most recent SCID therapies are based on gene therapy (GT) of hematopoietic stem cells (HSC), which are faced with many challenges. The new studies in the field of stem cells have made great progress in overcoming the challenges ahead. In 2006, Yamanaka et al. achieved "reprogramming" technology by introducing four transcription factors known as Yamanaka factors, which generate induced pluripotent stem cells (iPSC) from somatic cells. It is possible to apply iPSC-derived HSC for transplantation in patients with abnormality or loss of function in specific cells or damaged tissue, such as T-cells and NK-cells in the context of SCID. The iPSC-based HSC transplantation in SCID and other hereditary disorders needs gene correction before transplantation. Furthermore, iPSC technology has been introduced as a promising tool in cellular-molecular disease modeling and drug discovery. In this article, we review iPSC-based GT and modeling for SCID disease and novel approaches of iPSC application in SCID.

Objective(s): Hepatocellular carcinoma (HCC) is one of the leading fatal neoplasms and the most common primary liver malignancy worldwide. Peptide hormone ANGPTL8 (betatrophin) may act as an important regulator in HCC development through the Wnt/β-catenin pathway. We aimed to evaluate the effects of recombinant ANGPTL8 on Wnt/β-catenin signaling in human liver carcinoma cells (HepG2) and their viability. Materials and Methods: The expression of ANGPTL8 was conducted in the pET-21b-E. coli Bl21 (DE3) system and the produced peptide was purified. HepG2 cells were treated with different concentrations of ANGPTL8 (25, 50, 100, 150, 200, and 250 ng/ml) for 24, 48, and 72 hr. MTT assay was performed to detect the viability of treated cells, and apoptotic induction by ANGPTL8 was measured by flow cytometry assay. Finally, using qRT-PCR the mRNA levels of Wnt signaling modulators WIF-1 and β-catenin were evaluated in treated cells. Results: MTT assay showed that ANGPTL8 inhibits proliferation of HepG2 cells moderately in a time-independent manner. The highest concentration of the ANGPTL8, 250 ng/ml, reduced cell proliferation after 24, 48, and 72 hr similarly about 30%. In the same concentration of ANGPTL8, after 24 hr of treatment flow cytometry assay revealed a mild increase in early and late apoptosis up to 7.7 and 14.3%, respectively. The qRT-PCR showed that in a concentration-dependent and time-independent fashion, the expression of WIF-1 and β-catenin genes respectively increased and decreased significantly (PConclusion: Our findings suggest that ANGPTL8 may act as a moderate suppressor against HCC cell proliferation possibly via affecting Wnt signaling modulators.

Background: Cancer, a common disease in the world, is considered the second cause of mortality in developed countries. In the management of symptoms created by breast cancer (BC), pain is the most important. So, this study aimed to evaluate different dimensions of pain in patients using the McGill Pain Questionnaire. In this way, physicians could perform effective treatment for patients. Methods: This case study was done on BC patients aged 30-60 years old in some specialized cancer hospitals in Tehran. The utilized research instrument in this study was the McGill Pain Questionnaire. Data were analyzed by SPSS Software. Results: The BC pain in various dimensions as sensory, emotional, general understanding, and different pain types was studied on 166 women with BC. Our results indicated that pain was more in the sensory dimension in studied patients with an average rate of 80.4, which attributed 1 to 10 rows of the questionnaire. The most chosen words by BC patient in the sensory, emotional, and general understanding dimensions were as follow: the word “Sharp” with 69, “Troublesome” with 47, and “Nauseating” with 87 were the most frequent respectively. Conclusion: According to these results, it is possible to use effective and better treatment to reduce BC patients’ pain.

Background: Although the precise pathogenesis of acute lymphoblastic leukemia (ALL) remains unclear, studying gene-regulating mechanisms during ALL pathogeneses may shed light on the underlying mechanisms driving malignant behavior. There is some evidence showing the promoter hypermethylation and silencing of RASSF1A tumor suppressor gene in ALL cells; however, there is a lack of evidence for whether the gene indeed alters during different phases of ALL or in response to therapy. Thus, the current study aimed to clarify this issue using groups of adult ALL patients who have been scarcely investigated regarding expression levels and promoter methylation status. Materials and Methods: In this case/control study, the expression levels and methylation status of the gene promoter was evaluated using quantitative real-time PCR and methylation-specific PCR (MSP), respectively in adults with ALL. The study included peripheral blood of patients with newly diagnosed ALL (n=10), complete remission (CR) (n=10), or relapse (n=10), and 10 control samples from healthy individuals. Results: MSP results revealed an unmethylated status for almost all patients and control samples, except a case with relapsing ALL, which showed a hemimethylated pattern. RASSF1A also showed no difference in terms of gene expression in the patients compared with the control group (p>0.05). Conclusion: The results revealed an up-regulation of RASSF1A tumor suppressor in adult ALL patients experiencing CR, suggesting this to be a marker of therapy response. However, further investigations using more sensitive methylation detecting tools with larger sample sizes may better clarify the involvement of the promoter methylation of RASSF1A in these patients.

Recurrence in hepatocellular carcinoma (HCC) after conventional treatments is a crucial challenge. Despite the promising progress in advanced targeted therapies, HCC is the fourth leading cause of cancer death worldwide. Radionuclide therapy can potentially be a practical targeted approach to address this concern. Rhenium-188 (188Re) is a β-emitting radionuclide used in the clinic to induce apoptosis and inhibit cell proliferation. Although adherent cell cultures are efficient and reliable, appropriate cell-cell and cell-extracellular matrix (ECM) contact is still lacking. Thus, we herein aimed to assess 188Re as a potential therapeutic component for HCC in 2D and 3D models. The death rate in treated Huh7 and HepG2 lines was significantly higher than in untreated control groups using viability assay. After treatment with 188ReO4, Annexin/PI data indicated considerable apoptosis induction in HepG2 cells after 48 h but not Huh7 cells. Quantitative RT-PCR and western blotting data also showed increased apoptosis in response to 188ReO4 treatment. In Huh7 cells, exposure to an effective dose of 188ReO4 led to cell cycle arrest in the G2 phase. Moreover, colony formation assay confirmed post-exposure growth suppression in Huh7 and HepG2 cells. Then, the immunostaining displayed proliferation inhibition in the 188ReO4-treated cells on 3D scaffolds of liver ECM. The PI3-AKT signaling pathway was activated in 3D culture but not in 2D culture. In nude mice, Huh7 cells treated with an effective dose of 188ReO4 lost their tumor formation ability compared to the control group. These findings suggest that 188ReO4 can be a potential new therapeutic agent against HCC through induction of apoptosis and cell cycle arrest and inhibition of tumor formation. This approach can be effectively combined with antibodies and peptides for more selective and personalized therapy.

Background: About 8 percent of all cancers in human population are related to leukemia and it is one of the most common malignancies in children. The aim of this study was to compare the prevalence of age, gender and blood group types with the frequency of leukemia among the children with leukemia in Qazvin province during the 2006 to 2016.Materials and Methods: This was a cross-sectional analysis. Investigated population was 110 children and adolescents under 18 years in the hospitals of Qazvin province. The date collecting method was through review of medical records of the patients and their analysis performed by using SPSS version 16.Results: According to data from this study, leukemia ALL-L1 is more frequent in Qazvin than other types of leukemia, and children with ages 0-5 years was more than other age groups. This disorder is more common in boys than girls, and among the patients, the people who has A and O blood groups, and Rh + are the most abundant.Conclusion: such factors like age, gender and blood groups can use as prognostic factors in children leukemia. So that leukemia in children less than 5 years old is more than any other age. In addition to that; the incidence of leukemia ALL-L1 reduced with increasing age in the general population in Qazvin and number of boys with leukemia is more than girls.

Objective: Receptor activator of nuclear factor-kappa B ligand (RANKL) appears to be an osteoclast-activating factor, bearing an important role in the pathogenesis of multiple myeloma. Some studies demonstrated that U-266 myeloma cell line and primary myeloma cells expressed RANK and RANKL. It had been reported that the expression of myeloid and monocytoid markers was increased by co-culturing myeloma cells with hematopoietic stem cells (HSCs). This study also attempted to show the molecular mechanism of RANK and RANKL on differentiation capability of human cord blood HSC to osteoclast, as well as expression of calcitonin receptor (CTR) on cord blood HSC surface. Materials and Methods: In this experimental study, CD133+ hematopoietic stem cells were isolated from umbilical cord blood and cultured in the presence of macrophage colony-stimulating factor (M-CSF) and RANKL. Osteoclast differentiation was characterized by using tartrate-resistant acid phosphatase (TRAP) staining, giemsa staining, immunophenotyping, and reverse transcription-polymerase chain reaction (RT-PCR) assay for specific genes. Results: Hematopoietic stem cells expressed RANK before and after differentiation into osteoclast. Compared to control group, flow cytometric results showed an increased expression of RANK after differentiation. Expression of CTR mRNA showed TRAP reaction was positive in some differentiated cells, including osteoclast cells. Conclusion: Presence of RANKL and M-CSF in bone marrow could induce HSCs differentiation into osteoclast.

Objective: To determine the antimicrobial susceptibility patterns, the frequency of integrons and associated gene cassettes in Acinetobacter baumannii (A. baumannii) strains isolated from selected hospital intensive care units. Methods: During a ten-month period, 120 A. baumannii isolates were studied. The resistance rates to different classes of antimicrobial agents were determined. PCR was used to detect different types of integrons and associated gene cassettes. Results: The resistance rates to the majority of antibiotics tested were found to be between 39.3% and 99.1%. No isolate was observed to be resistant to colistin and polymyxin B. The rate of extensive drug-resistance among these clinical isolates was 62.5%. The prevalence of class 1 and 2 integrons was found to be 74.1% and 12.5%, respectively. Seven different gene cassettes (ampC, aacA4-catB8, ISAba1-blaOXA-23-GES-14, aadA2-cm1A6-GES-14-qacF, VIM-25-GES-24-qacF, dfrA5-ISAba1-blaOXA-51-blaOXA-40 and aadA2-GES-11-IMP-1) were observed in Class 1 integron-carrying strains. Three gene cassettes (IMP-4, VIM-2-VEB-aacA4 and dfrA2-sat-2-aadA4) were detected in class 2 integron-bearing A. baumannii strains. Conclusions: A high prevalence of integron was described among multidrug resistant A. baumannii in the hospital. The findings highlighted the need for continuous surveillance in order to prevent dissemination of multidrug resistance among A. baumannii strains in Iran.

Background and Aim: Different regulation processes have an effect on osteoblastic differentiation of mesenchymal stem cells (MSCs), and among them Wnt signaling pathway is particularly desirable. In Wnt signaling pathway, Adenomatous Polyposis Coli (APC) bind to β-catenin and induce its degradation, thereby acting as a negative regulator of canonical Wnt pathway. In this study, gene expression and DNA methylation of APC gene during osteoblastic differentiation were determined. Materials and Methods: In this experimental study, after the isolation of MSCs, the induction of osteoblastic differentiation was done. To confirm osteoblastic differentiation, alizarin red staining together with the expression of Alkaline Phosphatase (ALP) and osteocalcin as specific osteoblastic markers was performed. APC gene methylation status by MSP (Methylation Specific PCR) and gene expression status of APC gene using Real-Time PCR technique during different times were evaluated. Results: The results of alizarin red staining and the expression of ALP and osteocalcin confirmed osteoblastic differentiation. In addition, the results showed a significant decrease in the expression of APC gene on the 7th day of osteoblastic differentiation (P

Background: Prostate cancer (PC) is the second most common malignancy in men, accounting for 12.5% of total number of cancers. The development of molecular studies (such as transcriptomics analysis) helps to characterization of Cancer, development of new targets for therapy, and introduction of the novel prognostic and diagnostic biomarkers. Recent studies have confirmed Mammalian Sterile 20-Like kinase (MST1) as a tumor suppressor gene. In this study we focus on MST1 expression level in WBC of PC patients, due to inheritance pattern of PC. Material and Methods: This case-control study was conducted in two groups (20 patients with PC and 20 healthy individuals). After RNA extraction and cDNA synthesis, quantitative Real-Time PCR was done in order to determine the MST1 expression level. GAPDH was considered as internal control gene. Statistical analysis was performed by “Rotor-Gene Q series software 2.3.1” and “Rest 2.0.13 software”. Results: the study on 20 PC patients aged 50 to 70 years old and 20 healthy individuals shown that MST1 expression level in the WBC samples of PC patients have been reduced ≈62% compared to normal individuals. Conclusion: Introducing the reduced expression level of MST1 as a prostate cancer biomarker requires complementary research. However, in this study, biomarker validation and potential of MST1 has been approved

Background: Leukemia is the most common type of childhood cancer, with acute lymphoblastic leukemia being the most common subtype among children. Epigenetic factors, such as DNA methylation, are found to be important in leukemia. According to the National Cancer Institute, Wilms’ tumor (WT1) is among the most important tumor antigens. This study examines WT1 gene methylation as a diagnostic or therapeutic method in patients afflicted by ALL. Methods: 37 ALL patients were enrolled in this study before initiation of treatment. Twelve healthy subjects were used as the control group. After DNA extraction and conversion with bisulfite, WT1 gene methylation in samples was analyzed via the MSP technique. Results: The WT1 gene promoter was not methylated in patients or the control group. Conclusion: Due to the absence of WT1 gene promoter methylation in patients, as well as the healthy group, it is likely that methylation of the promoter of this gene is of no consequence in leukemia development, and subsequently cannot aid in its diagnosis.

Background: DNA methylation is an epigenetic mechanism that often modifies the function of genes and affects gene expression, which may sometimes lead to cancerdevelopment. Tumor suppressor genes such as VHL (Von Hippel-Lindau) may besilenced when affected by epigenetic alteration. This study investigated the DNAmethylation of VHL gene in ALL (Acute Lymphoblastic Leukemia) patients withmethylation specific PCR (MSP). Methods: The DNA of peripheral blood and bone marrow cells of 26 ALL patientsand 26 healthy control subjects were extracted, treated with bisulfite, and VHL genemethylation was subsequently analyzed using the MSP technique. Results: None of the patients showed signs of methylation in the VHL gene. Conclusion: Due to the lack of methylation in the VHL gene promoter in patientsand in the healthy control group, it is unlikely that the methylation of this gene canbe used in the diagnosis and prognosis of ALL patients.

Background: In spite of being vital to save the patients' life, blood transition may be dangerous and even fatal, too the aim of this study was to investigate the side effects (complications) of blood transfusion in the educational hospitals of Qazvin.Materials and Methods: This is a descriptive cross sectional and practical study that was carried out in 2010. In this study, all the patients of four health training centers in Qazvin, that have had blood transfusion and complications, were considered as a part of the statistical community. The instrument for data collection was checklist which was filled through an interview with blood bank manager and some other responsible individuals and scrutinizing files of patients who had blood injection among the blood products consumption, request for the packed cells was the most and for fresh frozen plasma was the least.Results: 75% of these people had only one blood injection and the maximum injection volume was 100cc which was done mostly in the evening. Most of the transfusion history belonged to 21-30 year olds in our statistical community. 56% of all Patients that had transfusion, possessed background of some disease such as heart problems (21.9%). More than half of them (2.56%) had a chill feeling complication transfusion and there was a significant relationship between the blood transfusion volume and itching complication.Conclusion: Existence of a continuous association between blood transfusion organization and hospitals is indispensable. Therefore, it seems that Hemovigilance system or computer connected network to send reports, between hospital centers and blood transfusion organization of Iran, can be an appropriate solution.

Background: Vast variety of intermediate factors including cell cycle regulators, growth factors, transcription factors, and signaling pathways are involved in hematopoietic stem cell (HSC) commitment and differentiation into distinct lineages. VHL, Ecad, and RUNX3 are among these. Epigenetics is currently introduced as a potential mechanism to control the gene regulation. The aim of this study is to reveal the correlation between the expression level and methylation pattern of mentioned genes after in vitro differentiation of cord blood HSCs into erythroid lineage mediated by erythropoietin.Materials and Methods: After isolation and expansion, the CD34+ cord blood stem cells were divided into two parts. The first part was used to extract the DNA and RNA and the second to differentiate into erythroid lineage. Methylation specific PCR (MSP) and Real-time PCR were used to determine the methylation status and expression levels of the genes, respectively.Results: Although the significant upregulation observed for VHL and Ecad genes and a down-regulation for RUNX3 gene after differentiation, no remarkable changes were seen in methylation pattern compared with cord blood HSCs by MSP technique.Conclusion: It is appearing that methylation pattern in promoter region has not an effective role in expression of VHL, Ecad, and RUNX3. Moreover, considering the inability of MSP method to detect subtle differences in methylation level a more sensitive method is needed to distinguish the methylation levels of these genes before and after erythroid differentiation.

Background: Several influential factors such as transcription factors and intracellular signaling components are involved in differentiation of stem cells into a specific lineage. P15 and p16 proteins are among these factors. Accumulating evidences has introduced the epigenetic as a master regulator of these factors during lineage specification. The main objective of this study is to determine the correlation between the expression level and methylation pattern of P15 and P16 genes in erythroid lineage after in vitro differentiation by erythropoietin (EPO).Materials and Methods: The purified and expanded CD34+ cord blood stem cells were differentiated into erythroid lineage in the presence of EPO. DNA was isolated from both cord blood stem cells and differentiated cells. The Real-Time PCR performed using cDNA and the isolated DNA was used in methylation Specific PCR (MSP) reaction for methylation pattern analysis in both pre and post differentiation stages.Results: The study demonstrated that P15 and P16 genes have partial methylation after erythroid differentiation by EPO. The Expression of P15 gene was higher after differentiation and the expression of P16 gene had a slightly decreased level in post differentiation stage.Conclusion: Significant increase in P15 gene expression after differentiation to erythroid lineage, suggests the remarkable efficacy of this gene in erythroid function. According to upregulation of P15 gene after differentiation despite unchanged methylation status and slight down regulation of P16 gene with slight hyper-methylation of the gene it can be suggested that although the methylation can affects the expression level of P16 gene, the P15 gene is not affected by this mechanism during erythroid differentiation mediated by EPO.

Background: Hematopoietic stem cell transplantation (HSCT) is a therapeutic approach in treatment of hematologic malignancies and incompatibility of bone marrow. Umbilical cord blood (UCB) known as an alternative for hematopoietic stem/ progenitor cells (HPSC) for in allogenic transplantation. The main hindrance in application of HPSC derived from umbilical cord blood is the low volume of collected samples. So, ex vivo expansion of HPSCs is the useful approach to overcome this restriction. Synthetic biomaterials such as nanofibers is used to produce synthetic niches. The aim of this study was the ex vivo expansion of hematopoietic stem cells on biocompatible nanofiber scaffolds. Methods: This study was done at Tarbiat Modares University from November 2012 to June 2013 and was a research study. Umbilical cord blood CD133+ hematopoietic stem cells were separated using MidiMacs (positive selection) system by means of monocolonal antibody (microbeads) CD133. Flow cytometry was used to assess the purity of cells. Cell culture was done on plate (2 Dimensional) and fibronectin conjougated polyether sulfone nanofiber scaffold (3 Dimensional). Colony assay test was used to asses the ability of colonization of cells. Results: Cell count analysis revealed the expansion of hematopoietic stem cells in cell culture plate (2D environment) and on nanofiber scaffold (3D environment) after 2 weeks. Expansion of cells in 2D environment was greater than 3D condition. Colony assay test revealed that the colonization ability of cells decreased after 2 weeks, but this decrease was lower in scaffold culture than plate culture. Conclusion: This study demonstrated that umbilical cord blood CD133+ hematopoietic stem cells can expand on fibronectin conjugated polyether sulfone scaffold and we can use this system for expanding of cells in vitro situation.

Background:The emergence and increase in the incidence of Extended-spectrum beta lactamase (ESBL) producing Escherichia coli has become an emerging challenge especially in hospitalized patients with UTI. The aim of the present study was to survey the frequency of bla CTX-M genotype in ESBL producing E. coli isolated from hospitalized patients with UTI and determination of their antibiotic resistance pattern.Material and methodsA total of 135 E. coli isolates were collected from isolated from patients with UTI. The isolates were subjected to confirmatory phenotype tests for the presence of ESBL. 75 E. coli isolates were confirmed as ESBL-positive by means of the Double disc synergy test. In vitro susceptibility of ESBL isolates to 15 antimicrobial agents amoxicillin, penicillin, ceftazidime, cefotaxime, cefoxitin, ceftriaxone, cefixime, cephalexin, co-trimoxazole, gentamicin, nalidixic acid, ciprofloxacin, nitrofourantoin, amikacin and imipenem was performed by Kirby-Bauer’s Disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI, 2012) guideline. PCR method was used to identify bla CTX-M gene in 75 ESBL positive strains.Results:PCR and sequence analysis showed that 75 (55.5%) isolates produced bla CTX-M genes. In vitro susceptibility of ESBL producing E. coli showed that all of them were resistant to amoxicillin and penicillin and The rates of resistance to the majority of tested antibiotics varied between 61% to 100 %, with the exception of amikacin (14.7%) and imipenem (2.7%). Our results showed that the frequency of bla CTX-M was strikingly high (93.3%).Conclusion:These data confirmed that the frequency of bla CTX-M genes were high among E. coli isolated from patients with UTI. The trend of multidrug-resistant profile has been associated with bla CTX-M gene is alarming. Therefore, it is very important to establish a routine screening of ESBL in clinical isolates to prevent dissemination of resistant isolates in health care settings.

Objective(s): Stem cell differentiation into different cell lineages depends upon several factors, cell cycle control elements and intracellular signaling elements, including P15INK4b and P16INK4a genes. Epigenetics may be regarded as a control mechanism which is affected by these factors with respect to their promoter structure. Materials and Methods: The CD34 + cord blood stem cells were purified, isolated and then expanded. The undifferentiated day genome was isolated from part of the cultured cells, and the seventh day differentiated genome was isolated from the other part after differentiation to erythroid lineage. The procedure was followed by a separate Real-Time PCR for the two genes using the obtained cDNA. The processed DNA of the former stages was used for MSP (Methylation Specific PCR) reaction. Finally, pre- and post differentiation results were compared. Results: After performing MSP for each gene, it became clear that P15INK4b gene has undergone methylation and expression in predifferentiation stage. In addition, its status has not been changed after differentiation. P15INK4b gene expression was reduced after the differentiation. The other gene, P16INK4a, showed no predifferentiation methylation. Itwas completely expressed methylated and underwent reduced expression after differentiation. Conclusion : Specific predifferentiation expression of P15INK4b and P16INK4a genes along with reduction in their expression after erythroid differentiation indicated animportant role for these two genes in biology of CD34+ cells in primary stages and before differentiation. In addition, both genes are capable of epigenetic modifications due to the structure of their promoters.

Background: Acute lymphoblastic leukemia (ALL) is a clonal malignant disorder characterized by an uncontrolled proliferation of immature T or B lymphocytes. Extensive studies have shown that the epigenetic changes, especially modified DNA methylation patterns in the regulatory regions through the DNA methyltransferase (DNMTs), play an important role in the development of genetic disorders and abnormal growth and maturation capacity of leukemic stem cells (LSCs).The aim of this study was to evaluate the changes in DNMT1 promoter methylation and its expression pattern in patients with ALL. Subjects and Methods: In this experimental study, methylation specific PCR (MSP) was used to assess the methylation status of DNMT1 promoter regions in samples collected from ALL patients (n=45) and healthy control subjects. According to this method, un-methylated cytosine nucleotides are converted to uracil by sodium bisulfite and the proliferation of methylated and un-methylated regions are performed using specific primers for target sequences. Results: None of the patients with B and T-ALL showed methylated promoter regions of the DNMT1 gene, while the methylation pattern of both pre-B ALL patients and the control group showed a relative promoter methylation. Conclusion: Analysis of promoter methylation patterns in various subgroups of ALL has revealed the importance of DNMT1 in the regulation of gene expression. Likewise, extensive data have also highlighted the methylation-based mechanisms exerted by DNAM1 as one of the main participants regulating gene expression in B-ALL and T-ALL patients. Investigation of the overall DNA methylation pattern offers significant improvements in the prediction of disease prognosis and treatment response.

Iron chelation therapy is used to reduce iron overload development due to its deposition in various organs such as liver and heart after regular transfusion. In this review, different iron chelators implicated in treatment of iron overload in various clinical conditions have been evaluated using more up-to-date studies focusing on these therapeutic agents. Deferoxamine, Deferiprone and Deferasirox are the most important specific US FDA-approved iron chelators. Each of these chelators has their own advantages and disadvantages, various target diseases, levels of deposited iron and clinical symptoms of the afflicted patients which may affect their selection as the best modality. Taken together, in many clinical disorders, choosing a standard chelator does not have an accurate index which requires further clarifications. The aim of this review is to introduce and compare the different iron chelators regarding their advantages and disadvantages, usage dose and specific applications.

Reprisal is a mechanism for the owner of a right to vindicate his right without the permission and consent of the debtor or usurper and without referring to the court when he does not have statutory evidence to offer to the courts. Valid evidence and proofs in the Shiite jurisprudence suggest the legitimacy of reprisal. Regulations and conditions of this institution of vindication of rights is raised in the jurisprudential sources and its limitations as well as its permissiveness are clarified there. With application of reprisal the creditor satisfies his right , the debtor is discharged, and the obligation is extinguished. Thus reprisal in legal studies can be raised under the title of extinction of obligations. Since reprisal is a legitimate institution applied in all societies and since its regulations have been dealt with in the Shiite jurisprudence it is necessary that the legislator declare reprisal as the seventh statutory cause of the extinction of obligations.

Objective: Thiopurine S-methyltransferase (TPMT) catalyses the S-methylation of thiopurinedrugs. Low activity phenotypes are correlated with several mutations in the TPMTgene and adverse drug reactions. The molecular basis for dissimilar enzymatic activityof TPMT has been established in Caucasians, African-Americans and Southwest Asians,but it remains to be elucidated in Iranian population. Until present, no study on Iranianpopulation has been performed on the known alleles of TPMT. The aim of this study wasto investigate the frequencies of four of the most common variants of this gene.Materials and Methods: This study was conducted during 2007 at the Department of Hematology,Tarbiat Modares University, Tehran, Iran. Using PCR-RFLP and allele specificPCR techniques, allelic variants of the TPMT gene TPMT*2(G238C), TPMT*3B (G460A),TPMT*3C (A719G) and TPMT*3A (G460A and A719G) were genotyped in a normal populationof 127 Iranians.Results: In this study TPMT*2 showed a prevalence of 7.08%. TPMT*3C and *3A werefound in 2.47% and 2.18% of the samples, respectively. TPMT*3B variant was not detectedin Iranian subjects. 112 out of 127 participants showed homozygote wild type allele.Conclusion: This study is the first to analyze TPMT allele frequencies in a sample ofIranian population and indicates that TPMT*2 is the most common allele (7.08%) in thispopulation. These results can help to organize national pretreatment strategies in patientswith acute lympho blastic leukemia (ALL) or other diseases requiring thiopurine medicationin their standard therapy.

Background: Early diagnosis and treatment of leukemia patients remains a fundamental aim in clinical oncology, especially in developing country. Present study highlights the basic requirements of these patients in Iran. Better understanding of these issues may lead to improve the healthcare standards toward leukemia diagnosis and treatment. Methods: This descriptive study included 101 specialists in hematology-oncology and pathology serving in oncology centers. The participants were then asked to fill out a standard questionnaire on the issues around diagnosis and treatment of blood malignancies. Results: According to specialists, unfair distribution of facilities across the country, delayed diagnosis of disease, absence of psychological support for patients, and insufficient financial support were the main reasons of inappropriate diagnosis and treatment in leukemia patients. Conclusions: Our results show that making an amendment to health policies by preparing well-equipped medical centers in all provinces, improving the morale of patients through consultation during the process of treatment, and above all, subsiding leukemia patients' financial problems will promote the health standard regarding the leukemia diagnosis and treatment in Iran.

Objective. Plasmid-mediated quinolone resistance (PMQR) plays an important role in the development of clinical resistance to quinolone. The aim of this study was to investigate PMQR determinants among extended-spectrum β-lactamases- (ESBL-) producing Klebsiella pneumoniae recovered from patients with nosocomial urinary tract infection (UTI). Methods. A total of 247 ESBL-producing K. pneumoniae isolates were collected from 750 patients with UTI. ESBL production was confirmed by double disc synergy test and combined disc diffusion test. The prevalence of PMQR determinants among ESBL-producing K. pneumoniae was assessed using PCR method. Results. The rates of resistance to antimicrobial agents in present study varied from 14.2% to 98.8%. In comparison with other PMQR genotypes, the frequency of aac(6′)-Ib (68.8%) was strikingly high. Of the 247 isolates tested, qnrA, qnrB, qnrS, and qepA genes were present in 3.6%, 1.6%, 1.2, and 2%, respectively. oqxA and oqxB were detected in 56.7% and 54.6% of isolates. The predominant coexisting ESBL and PMQR profile among our isolates included blaCTX-M and aac(6′)-Ib, oqxA, oqxB (28.3%) and blaTEM, blaSHV and aac(6′)-Ib, oqxA, and oqxB (19.4%) profile. Conclusion. Given the linkage observed between resistance to quinolones and beta lactam antibiotics, therapeutic protocol with fluoroquinolones and beta lactam antibiotics should be seriously revised in Tehran hospitals.